RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-505
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Last modified: 9 February 2013, 16:03
*RMgm-505| Successful modification | The gene/parasite could not be changed/generated by the genetic modification. |
| The following genetic modifications were attempted | Gene disruption |
| Number of attempts to introduce the genetic modification | 3 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 22184632 Reference 2 (PMID number) : 15003842 |
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| Parent parasite used to introduce the genetic modification | |
| Rodent Malaria Parasite | P. berghei |
| Parent strain/line | P. berghei ANKA |
| Name parent line/clone | P. berghei ANKA 1037m1f1mocl1 (RMgm-32) |
| Other information parent line | P. berghei ANKA 1037m1f1mocl1 (1037cl1; RMgm-32) is a reference ANKA mutant line which expresses GFP-luciferase under control of a schizont-specific promoter (PubMed: PMID: 20019192). It does not contain a drug-selectable marker and has been selected by FACS sorting based on GFP expression |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | J. Fonager, E.M. Pasini, C.J. Janse, B.M Franke-Fayard |
| Name Group/Department | Leiden Malaria Research Group |
| Name Institute | Leiden University Medical Center (LUMC) |
| City | Leiden |
| Country | The Netherlands |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1433600 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1218000 | ||||||||||||||||||||||||
| Gene product | thrombospondin-related apical membrane protein | ||||||||||||||||||||||||
| Gene product: Alternative name | PTRAMP | ||||||||||||||||||||||||
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| Details of the genetic modification | |||||||||||||||||||||||||
| Inducable system used | No | ||||||||||||||||||||||||
| Additional remarks inducable system | |||||||||||||||||||||||||
| Type of plasmid/construct used | Plasmid double cross-over | ||||||||||||||||||||||||
| PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
| Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
| Plasmid/construct map | |||||||||||||||||||||||||
| Plasmid/construct sequence | |||||||||||||||||||||||||
| Restriction sites to linearize plasmid | Asp718I/EcoRI | ||||||||||||||||||||||||
| Partial or complete disruption of the gene | Unknown | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | |||||||||||||||||||||||||
| Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
| Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | Name of unsuccessful attempts: exp. 1250/1306/1486 The gene has been targeted for disruption/deletion using a construct for targeted gene disruption by double cross-over homologous recombination. The primers of the two target sequences of the gene are provided below. The unsuccessful attempts to disrupt the gene might indicate an essential function during asexual blood stage development. The gene has been selected for targeted gene disruption based on its presence in a proteome of membranes of infected red blood cells. See also Thompson et al., 2004. Mol. Biochem Parasitol. 134: 225-32: PTRAMP; a conserved Plasmodium thrombospondin-related apical merozoite protein. In this study the protein has been localised in the micronemes of P. berghei merozoites. However, The P. falciparum ortholog (PfTRAMP) localizes in the rhoptries. See Siddiqui et al, 2013 Cell Microbiol). | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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